Abstract
BackgroundWe aimed to evaluate the potential enhancing effect of celastrol on the stemness of human tendon-derived stem cells (hTSCs) in vitro and the underlying molecular mechanisms.MethodsThe capability of hTSC self-renewal was assessed by cell proliferation and colony formation as determined with the CCK-8 kit. Adipogenesis, chondrogenesis, and osteogenesis were determined by Oil Red O, Alcian Blue, and Alizarin Red staining, respectively. The relative mRNA levels of Sox9, PPARγ, Runx2, Smad7, and HIF1α were determined by real-time polymerase chain reaction (PCR). The levels of Smad7 and HIF1α protein were measured by immunoblotting. The chromatin immunoprecipitation (ChIP) assay was used to assess the direct binding of HIF1α to the Smad7 promoter. Suppression of Smad7 induced by hypoxia was examined using the luciferase reporter assay.ResultsWe found that treatment with celastrol resulted in improvement in both the multi-differentiation potential and self-renewal capability of hTSCs. Celastrol elicited hypoxia and subsequently suppressed the expression of Smad7 through direct association with the hypoxia response element consensus sequence. Further, we demonstrated that both Smad7 and HIF1α were involved in the beneficial effects of celastrol on the differentiation and self-renewal of hTSCs.ConclusionsWe demonstrated the positive effect of celastrol on the stemness of hTSCs and elucidated the essential role of the HIF1α-Smad7 pathway in this process.
Highlights
We aimed to evaluate the potential enhancing effect of celastrol on the stemness of human tendonderived stem cells in vitro and the underlying molecular mechanisms
Celastrol improves the self-renewal ability of Human tendon-derived stem cell (hTSC) Accumulating evidence supports the crucial role of hypoxia in the proliferation and differentiation of TSCs
We first treated hTSCs with different doses of celastrol (0, 1, 2, and 4 μM) for 24 h, and evaluated their self-renewal capacity based on both proliferation and colony formation assays
Summary
We aimed to evaluate the potential enhancing effect of celastrol on the stemness of human tendonderived stem cells (hTSCs) in vitro and the underlying molecular mechanisms. A growing body of evidence supports the potential therapeutic value of hTSCs in regeneration of the tendon, and intensive investigations have focused on the characterization, Wu et al Stem Cell Research & Therapy (2017) 8:274 isolation, expansion, and differentiation of hTSCs in vitro [6,7,8]. These findings from basic biological research are being proactively translated into clinical applications [9]. Due to the critical role of hypoxia in hTSC biology, an agent that is able to induce hypoxia in hTSCs is likely to contribute to the proliferation and differentiation of the stem cell culture and potentially to subsequent stem cell therapy for tendon repair
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